Fusion rp column

Manufactured by Phenomenex

Sourced in United States

The Fusion-RP column is a reversed-phase liquid chromatography column designed for the separation and analysis of a wide range of organic compounds. It features a proprietary stationary phase that provides efficient and reproducible separations.

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Lab products found in correlation

G6470a triple quad system, by Agilent Technologies (2 mentions) 1290 infinity 2, by Agilent Technologies (2 mentions) Microtof q 2, by Bruker (1 mentions)

Spelling variants of this product

Synergi Fusion-RP column (20 citations) Synergi Fusion-RP C18 column (12 citations) Fusion-RP C18 column (5 citations) Synergy Fusion RP column (3 citations) Synergi Fusion RP-18 column (3 citations) Synergi Fusion-RP 80A column (3 citations) C18 Synergi Fusion-RP 80i columns (2 citations) Fusion-RP 80A column (2 citations) Fusion RP−18 column (2 citations) Synergi 2.5 mM Fusion-RP 100A° (10033.0 mm) LC column (2 citations)

5 protocols using fusion rp column

HPLC Analysis of Anthocyanins

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HPLC analysis of anthocyanins was performed according to the method described by Nielsen et al. [46 (link)] with some modifications. In short, 5 µL of the eluate was analyzed using an Agilent HPLC Model HP 1200 equipped with a diode array detector (DAD). Separation was performed on a Phenomenex^®Fusion RP column (250 mm × 4.6 mm; particle size = 4 µm) using a mobile phase consisting of water/ formic acid (95:5 v/v) (A) and acetonitrile (B). Elution profile: 0–16 min, 3%–9% B; 16–30 min, 12% B; 30–54 min, 33% B; 54–58 min, 90% B; 58–62 min, isocratic 90% B. The anthocyanins in the eluate were detected at 520 nm and a temperature of 25 °C. Their amounts were quantified by calibration with the standards of cyanidin-3-O-glucoside and cyanidin-3-O-rutinoside and expressed in mg·100 g⁻¹ dry mass (DM). An example chromatogram of anthocyanins separation is shown in Supplementary Material (Figure S2).
The lyophilized powder (50 mg) was extracted in 2 mL of 60% methanol acidified with 1% formic acid for 20 min in an ultrasonic bath. The suspension was then centrifuged (7000× g, 10 min). The resulting extract for phenolic compound determination was filtered through a PTFE filter (0.45 µm, 15 mm).

Wojtania A, & Mieszczakowska-Frąc M. (2021). In Vitro Propagation Method for Production of Phenolic-Rich Planting Material of Culinary Rhubarb ‘Malinowy’. Plants, 10(9), 1768.

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Quantitative LCMS Analysis of Compounds

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For quantification, an Agilent 1290 Infinity II equipped with a DAD combined with an Agilent Technologies G6470A Triple Quad system and electrospray ionization (ESI-MS, Agilent Jetstream) was used. Operating parameters were as follows: positive-ion mode; skimmer voltage, 15 V; cell accelerator voltage, 5 V; N₂ gas temperature, 230°C; N₂ gas flow, 6 liters/min; sheath gas (N₂) temperature, 400°C with a flow of 12 liters/min; capillary voltage, 2500 V; nozzle voltage, 0 V; and nebulizer, 40 psi. The instrument was operated in dynamic MRM mode with the method listed in table S2. The mobile phases were as follows: A as 5 mM NH₄OAc aqueous buffer, brought to pH 5.6 with glacial acetic acid, and B as pure acetonitrile. A Synergi Fusion-RP column (Phenomenex, Torrance, CA, USA; Synergi 2.5 μm Fusion-RP 100 Å, 150 × 2.0 mm) at 35°C and a flow rate of 0.35 ml/min was used. The gradient began with 100% A for 1 min and increased to 10% B by 5 min and to 40% B by 7 min. The column was flushed with 40% B for 1 min and returned to starting conditions to 100% A by 8.5 min followed by re-equilibration at 100% A for an additional 2.5 min. For quantification, a stable isotope-labeled internal standard (46 ) was added to the samples before injection. Data analysis was performed as previously described (46 ).

Ignatova V.V., Kaiser S., Ho J.S., Bing X., Stolz P., Tan Y.X., Lee C.L., Gay F.P., Lastres P.R., Gerlini R., Rathkolb B., Aguilar-Pimentel A., Sanz-Moreno A., Klein-Rodewald T., Calzada-Wack J., Ibragimov E., Valenta M., Lukauskas S., Pavesi A., Marschall S., Leuchtenberger S., Fuchs H., Gailus-Durner V., de Angelis M.H., Bultmann S., Rando O.J., Guccione E., Kellner S.M, & Schneider R. (2020). METTL6 is a tRNA m3C methyltransferase that regulates pluripotency and tumor cell growth. Science Advances, 6(35), eaaz4551.

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Quantitative Analysis of Metabolites by LC-MS/MS

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For quantification, an Agilent 1290 Infinity II equipped with a DAD combined with an Agilent Technologies G6470A Triple Quad system and electrospray ionization (ESI-MS, Agilent Jetstream) was used. Operating parameters were as follows: positive ion mode, skimmer voltage 15 V, cell accelerator voltage 5 V, N₂ gas temperature 230°C and N₂ gas flow 6 L/min, sheath gas (N₂) temperature 400°C with a flow of 12 L/min, capillary voltage of 2500 V, nozzle voltage of 0 V, and the nebulizer at 40 psi. The instrument was operated in dynamic MRM mode with the method listed in Supplemental Table S2b. The mobile phases were A as 5 mM NH₄OAc aqueous buffer, brought to pH 5.6 with glacial acetic acid and B as pure acetonitrile. A Synergi Fusion-RP column (Phenomenex, Synergi 2.5 µm Fusion-RP 100 Å, 150 × 2.0 mm) at 35°C and a flow rate of 0.35 mL/min was used. The gradient began with 100% A for 1 min, increased to 10% B by 5 min, and to 40% B by 7 min. The column was flushed with 40% B for 1 min and returned to starting conditions to 100% A by 8.5 min, followed by re-equilibration at 100% A for an additional 2.5 min.

Ignatova V.V., Stolz P., Kaiser S., Gustafsson T.H., Lastres P.R., Sanz-Moreno A., Cho Y.L., Amarie O.V., Aguilar-Pimentel A., Klein-Rodewald T., Calzada-Wack J., Becker L., Marschall S., Kraiger M., Garrett L., Seisenberger C., Hölter S.M., Borland K., Van De Logt E., Jansen P.W., Baltissen M.P., Valenta M., Vermeulen M., Wurst W., Gailus-Durner V., Fuchs H., Hrabe de Angelis M., Rando O.J., Kellner S.M., Bultmann S, & Schneider R. (2020). The rRNA m6A methyltransferase METTL5 is involved in pluripotency and developmental programs. Genes & Development, 34(9-10), 715-729.

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Quantitative Bioanalysis of Imatinib and its Metabolites in Plasma

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Calibration standards and QC samples in plasma were prepared as follow: 3 μL of standard or QC working solutions were added to 297 μL of pooled plasma and vortexed for 10 s. Five microliters of spiked or patients’ plasma sample were precipitated with 245 μL of the extraction solution (10 ng/mL of IMA-D8). The mixture was vortexed for 10 s and then centrifuged for 10 min at 16000 g and 4°C. After that, 200 μL of the supernatant were transferred to a polypropylene autosampler vial for the subsequent analysis. The quantification was conducted with a LC-MS/MS method in-house developed and validated. The chromatographic separation was obtained with a Synergi Fusion-RP column (4 μm, 80 Å, 50 x 2.0 mm) coupled with a Fusion-RP security-guard pre-column (4 x 2.0 mm) (Phenomenex) under gradient condition using 0.1% formic acid/bidistilled water (v/v) and methanol/isopropanol (9:1, v/v) with 0.1% formic acid (v/v). The quantification was conducted using the following quantifier transitions: IMA m/z 494.4 > 394.2, norIMA m/z 480.4 > 394.2 and IS m/z 502.4 > 394.2. The method was linear over the concentrations range of 30–7500 ng/mL for IMA and 6–1500 ng/mL for norIMA.

Iacuzzi V., Posocco B., Zanchetta M., Montico M., Marangon E., Poetto A.S., Buzzo M., Gagno S., Buonadonna A., Guardascione M., Casetta B, & Toffoli G. (2019). Development and validation of LC-MS/MS method for imatinib and norimatinib monitoring by finger-prick DBS in gastrointestinal stromal tumor patients. PLoS ONE, 14(11), e0225225.

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LC-MS Analysis of Fractional Samples

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The fractions obtained were analysed by liquid chromatography mass spectrometry (LC-MS). Analyses were carried out on a Shimadzu Prominence Liquid Chromatography system coupled to a quadrupole time-of-flight mass spectrometer (Micro-TOFQII; Bruker Daltonics, Billerica, MA, USA) with an ESI as the ion source. Separations were achieved using a Fusion-RP column (150 mm × 2 mm, 4 μm; Phenomenex^®) protected with a guard column of the same material. Samples were eluted using a mobile phase A (water, 0.1% formic acid and 5 mM ammonia formate) and a mobile phase B (acetonitrile), at a flow rate of 0.2 mL/min. The ionization source conditions were as follows: positive ionization; capillary potential of 4000 V; temperature and flow of drying gas (nitrogen) of 5 L/min and 200 °C, respectively; nebulizer pressure of 35 psi. Mass spectra were acquired using full scan mode over the range of m/z from 50 to 1600.

Paiva F.C., Ferreira G.M., Trossini G.H, & Pinto E. (2017). Identification, In Vitro Testing and Molecular Docking Studies of Microginins’ Mechanism of Angiotensin-Converting Enzyme Inhibition. Molecules : A Journal of Synthetic Chemistry and Natural Product Chemistry, 22(12), 1884.

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